Carotenoid formulation

ABSTRACT

The present invention discloses the use of a lycopene coated with a water non-soluble thin film comprising amphiphilic protein polymer for coloring with red color, foods, pharmaceuticals or cosmetics having fat and/or oil contents higher than 5%. The invention further discloses a process for the preparation of stable lycopene formulation comprising: (a) treating an isolated protein to form a protein in a molecular form; (b) dispersing lycopene in an aqueous solution comprising an isolated protein in a molecular form; (c) grinding said dispersion to form lycopene particle size of 1 to 10 μm forming an homogenized mixture comprising line particles; and optionally (d) drying the homogenized mixture.

FIELD OF THE INVENTION

The present invention is in the field of lycopene formulations, aprocess for their preparation and their use as additives and inparticular as colorants retaining lycopene's unique color.

BACKGROUND OF THE INVENTION

Lycopene, belonging to the carotenoid family of natural substances isabundant in various fruits and vegetables. The compound as found innature has a trans/cis ratio of 90:10, is insoluble in water and has arather limited solubility in oil or fat. Studies have shown thatfrequent consumption of lycopene is associated with decrease of chronicdisorders such as cardiac and circulatory disorders. In addition, it maycontribute to prevention of several types of cancers. Its protectivefunction is attributed to the fact that lycopene acts as an effectiveantioxidant. Its unique intense red color led to its use as a naturalfood colorant. Lycopene may be obtained, either synthetically (EP382,067) or extracted from tomatoes (WO 97/48,287) which have arelatively high concentration of lycopene.

However, use of lycopene as a food supplement is associated with severalproblems. Its solubility profile is problematic, being insoluble inwater and only sparingly soluble in oils and fats. In addition, it iseasily oxidized. In addition, lycopene exhibits its unique red coloronly as long as it is dispersed in water and the particles have a sizeof 1-10 μm, preferably 1-3 μm. Particles of higher dimensions are lesseffective colorants. Furthermore, upon contact with oil the lycopeneparticles dissolve imparting an orange to yellow color.

As a result, lycopene is used only in dedicated formulations, where thelycopene is protected from oxidation by certain additives and/or thelycopene is present as finely divided particles. WO 91/06292 and WO94/19411 describe the grinding of β-carotene achieving a particle sizeof ca. 2-10 μm. Dry powders containing lycopene for use as waterdispersible powders were reported in EP 832569 and WO 98/16204. Stablepulverulent lycopene formulations comprising lycopene having a degree ofcrystallinity of greater than 20% are disclosed in U.S. Pat. No.6,235,315. The stability of the pulverulent formulation is augmented byaddition of protective colloids, stabilizers and plasticizers. U.S.2002/0128325 discloses a process for producing powders of stabilizedcarotenoids, where a carotenoid is dispersed in a solution containing aprotective colloid and lactose, and the use of such powders inpharmaceuticals, food and cosmetics.

SUMMARY OF THE INVENTION

The present invention is based on coating solid crystallized micronizedlycopene particles with a thin film comprising of water solubleamphiphilic biopolymers such as proteins or a mixture of proteins andhydrocolloids to form, once precipitated on the lycopene, a film ofwater non-soluble net work. Such film formation yields protectedlycopene that once placed in an aqueous phase containing lipids,prevents the lycopene from migration in its solid form or molecular forminto the lipids/oil/fats. Such coated lycopene thus makes it possible touse lycopene in lipid phases maintaining its typical red color, wheremigration into the lipid/oil/fat phase would result in a molecularyellow hue. Such a coated lycopene may thus be used as an additive orcolorant in foods, pharmaceuticals or cosmetic preparations having lipidcontents higher than 5%.

Thus the present invention is directed to the use of a lycopene coatedwith a water non-soluble thin film comprising amphiphilic proteinpolymer for coloring with red color, foods, pharmaceuticals or cosmeticshaving fat and/or oil contents higher than 5%. The water non-solublefilm may further comprise a colloid. The coated lycopene can be of anysource of lycopene, e.g., synthetic lycopene, tomato pulp, or lycopeneextracted from biomass, preferably from tomato pulp.

The present invention is further directed to a process for thepreparation of stable lycopene formulation comprising:

-   -   (a) treating an isolated protein to form a protein in a        molecular form;    -   (b) dispersing lycopene in an aqueous solution comprising an        isolated protein in a molecular form;    -   (c) grinding said dispersion to form lycopene particle size of 1        to 10 μm forming an homogenized mixture comprising fine        particles; and optionally    -   (d) drying the homogenized mixture.

The process may further comprise the step of mixing said lycopeneparticles with at least one colloid in water followed by drying.Optionally, the lycopene may be grinded separately and then added to theaqueous solution of isolated protein.

The lycopene used may be of any source, e.g., synthetic, naturallyextracted or a crude biomass-comprising lycopene. The formulation mayfurther comprise antioxidants and/or emulsifiers. All additives to thelycopene forming the formulation are food-grade.

The present invention is further directed to a lycopene containing drypowder obtained by the above process. Such a dry powder is havingtypically a lycopene content of 1% to 15% (w/w), preferably from 4% to8%.

The present invention is yet further directed to a method for coloringin red natural color, foods, cosmetics or pharmaceuticals having an oilor fat contents of at least 5% using the dried homogenized lycopeneformulation.

DETAILED DESCRIPTION OF THE INVENTION

The following description is illustrative of embodiments of theinvention. The following description is not to be construed as limiting,it being understood that the skilled person may carry out many obviousvariations to the process.

Throughout the description, percentages are weight by weight unlessspecifically noted differently. The term “lipid” encompasses oil andfat.

As mentioned the present invention is directed to the use of lycopenecoated with a water non-soluble thin film comprising amphiphilic proteinpolymer for coloring with red color, foods, pharmaceuticals or cosmeticshaving lipid contents higher than 5%. The invention is further directedto a process for formulating lycopene enabling its use in foods,pharmaceutical or cosmetic preparations having a lipid content higherthan 5% maintaining its red color. Lycopene upon coming into contactwith foods pharmaceutical or cosmetic preparations having such oil orfat contents tends to dissolve in the lipid phase. The result of suchdissolving is the loss of its unique red color. Thus in order to preventsuch migration and enable the use of lycopene as an additive to suchmatrices having high oil or fat contents maintaining its unique redcolor, a process for the coating of lycopene is disclosed. According tothe present invention the lycopene is coated with a transparent filmthus prohibiting the migration of the lycopene into the oil or fatphases. The transparent film is composed of an amphiphilic protein.Another option is to add a colloid to the protein-coated lycopene. Thelycopene that may be coated according to the present invention may be inany of its forms, non-limiting examples being synthetic lycopene,lycopene extracted from its natural source, e.g. tomatoes or even acrude biomass-comprising lycopene. The amphiphilic proteins used by thepresent invention are proteins which posses lipophilic amino acids intheir chains. Examples of amino acids that impart the desiredlipophilicity are leucine, isoleucine, phenylalanine and valine. Inaddition, proteins that consist of low contents of cystine and cysteinin their chains and that tend to cross-link only under extremeconditions of pressure or temperature can be used. Such proteins aresurface active and upon an interaction with lycopene, in any of itsforms, interact with lycopene fibers and adsorb thereon, forming a thinlayer of protein coating the lycopene fiber. Non limiting examples ofproteins are hydrophobically modified soy proteins, whey proteinisolates, egg albumin, lysozymes, modified pea proteins and gelatin ormixtures thereof. Modification of proteins may be done either byhydrolizing the proteins, by chemical reaction or by enzymes. Theprotein in its natural 3-dimensional structure can not be used as acoating film. Rather, the protein should initially be transferred intomolecular form by dispersing the protein in water, adjusting the pH tothe range of 9 to 10, heat the dispersion, cool and lyophilize.

Certain colloids may be attracted to proteins by hydrophobic or bycharge interactions to form protein-colloid complexes. Thus in additionto coating of lycopene by amphiphilic protein, the lycopene according tothe present invention may be coated by a protein-colloid film resultingfrom the interaction of the amphiphilic protein with colloids, forming acoating film comprising of protein and colloid. Such a film, if stronglyand effectively adsorbed, is resistant to water dilution and protectslycopene from migrating into oil or fat phases. Non limiting examples ofcolloids are proteinceous polysaccharides hydrocolloid or protectivecolloids, wherein said proteinceous polysaccharides hydrocolloid isselected from the group consisting of gum arabic, xanthan gum, amidatedstarch, amidated pectins; and said protective colloids are selected fromthe group consisting of food grade polysaccharides, polysaccharides, orgums selected from pectins, alginates, xanthan, tragacanth, or theirmodified structures, modified starch, moditified chitosans,maltodextrin, modified methylcellulose, galactomannan or mixturesthereof.

The lycopene powder, either as a dry biomass, crystalline lycopene orwet biomass, is formulated according to the present invention bydispersing lycopene in an aqueous solution comprising an isolatedprotein in molecular form and the dispersion is grind homogenized togive lycopene particles having a particle size of from 1 to 10 μm,preferably from 1 to 5 μm and most preferably from 1 to 3 μm. It shouldbe understood that the grinding is a prerequisite enabling the coatingof the lycopene by the protein. Such grinding ensures the coating of theentire lycopene fiber in a protein film. The resulting solutioncomprising of the coated lycopene may be used for coloring and/oradditive in foods, pharmaceuticals or cosmetics. Alternatively, thelycopene may be dried to yield a powder, which has a lycopene content offrom 1% to 15% (w/w), preferably from 2% to 10% and most preferably from4% to 8%.

As mentioned the protein film coating the lycopene may further interactwith colloids utilizing possible hydrophobic or charge interactions thusforming a coating film comprising of colloid bound protein. It should benoted that the coating film, either protein or protein/colloid mayfurther comprise antioxidants and/or emulsifiers added at the stage ofgrinding. Such antioxidants and/or emulsifiers are added at the stage ofadded at least one colloid. Non limiting examples of antioxidants areascorbic acid, citric acid, tocopherol or ascorbel palmitate. A mixtureof ascorbic and citric acids (1:1 wt. ratio) is added to the mixturecomprising of the lycopene/protein dry powder and colloid in water. Nonlimiting examples of emulsifiers are Tween, polyglycerols esters, sugaresters, lecitins, castor oil and ethoxylated castor oil.

According to a specific embodiment of the present invention, the coatedlycopene is used as a colorant for foods, pharmaceuticals and cosmetics.Accordingly, the coated lycopene formulation is added to foods,pharmaceuticals and cosmetics to impart a red color. Said lycopeneformulation is added to the foods, pharmaceuticals and cosmetics duringthe process of their preparation. The stage at which the lycopeneformulation is added in said preparation process may vary and may bedetermined by the skilled artisan.

The amount of lycopene formulation effective for imparting the desirablered color to the product may vary. The amount of lycopene formulationadded is calculated on the basis of lycopene, wherein the amount oflycopene is 10 to 200 ppm.

The invention will now be described by the following non-limitingexamples, where it should be stressed that calculation of tomato pulp tosoy mass weight ratios is based on 10-200 ppm Lycopene in said finalcomposition. The tomato pulp is a biomass of tomato after the removal ofmost of the water-soluble fraction. The tomato pulp may be used as is ordried prior to its use.

EXAMPLES Example 1 Process for Coating Crystalline Lycopene

100 g isolated Soy protein SUPRO® EX34K (Protein TechnologiesInternational Belgium) were dissolved in 1000 g water (30 minutes at40-50° C.). 50 g pure crystalline lycopene (Lyc-O-Mato® 70%) were addedto the solution. Thorough mixing resulted in a homogenous mixture, whichwas grounded in a ball-mill to yield lycopene having a particle size of1-3 μm. The ground lycopene was mixed with 650 g modified starch (Miracup, Staley, USA) and 1000 g water (TDS of mixture ca. 30%). The mixturewas immediately spray dried to a cold water dispersible oil resistantpowder having a lycopene contents of ca. 6% (Lyc-O-Mato® 6% OR).

Example 2 Industrialized Process for Coating Crystalline Lycopene

2.5 kg of Soy Isolated Protein EX 33K (P Production) were dissolved toyield a 6% protein water solution (41.7 kg solution) pH adjusted in therange of 9-10 and the solution was cooked for 1 hr at 80° C. Theantioxidants, Ascorbyl Palmitate and α-Tocopherol were added at anamount of 1% (0.04 kg each) from total solids (pH maintained in therange of 9-10). 1 kgp of lycopene (1.54 kg 65% lycopene) werehomogenized with the protein solution {where the resulting 43 kgsolution may be used as a mixture for drinking comprising of 4.12 kgtotal solids and 2.32% lycopene}. The homogenized protein-lycopenesolution is ground after which are add 7.7 kg of Mira cap (modifiedstarch) and homogenized. TDS before drying 23.3% Lycopene concentrationin dry finish product 8.04% with 5% moisture. The quantities of eachcomponent are summarized in the table below. Lyc-O-Mato 70% 1 kg Pure(1.54 kg as is) Protein EX 33K 2.5 kg Ascorbyl Palmitate 0.04 kgα-Tocopherol 0.04 kg Mira cap 7.7 kg Total solids 11.82 kg Lycopene indry product 8.46% Lycopene in dry powder with 8.04% 5% moisture

Application: 1000 g of vegetable protein mass (fat contents of 16%)mixture with 0.8 g of Lyc-o-mato®8% OR. Pack in polyethylen casing andcooking 30 min at 90° C. Colour after cooking—pink, without orange hue.

Example 3

Disperse 0.1 g isolated soy protein EX-34K in 99 g H₂O. Adjust pH ofsuspension to 9.2 with 0.5 M NaOH. Heat the dispersion at 70° C. for 30min. Cool to room temperature and dry by lyophilization. Dissolve 0.1 gof lyophilized powder (water-soluble protein) in 100 g H₂O. Add 10 g oftomato pulp (Lycored). Homogenize pulp suspension using “Silverson”dispersing machine (1000 rpm). Dry pulp with protein by spray drymethod.

Application: Add dried powder (10.1 g) to 1270 g soy mass (“Tivall”),homogenize, pack under vacuum in special cover, heat in water (90° C.,15 min.) and store. The soy mass remains with a pink-red color.

Example 4

Disperse 0.1 g isolated soy protein EX-34K in 99 g H₂O. Adjust pH ofsuspension to 9.2 with 0.5 M NaOH. Heat the dispersion at 70° C. for 30min. and cool to room temperature. Add 10 g of tomato pulp (Lycored) toprepared solution (of water-soluble protein). Continue further like inExample 3.

Example 5

Disperse 0.05 g isolated soy protein EX-34K in 99 g H₂O. Adjust pH ofsuspension to 10.2 with 0.5 M NaOH. Heat the dispersion at 70° C. for 30min and cool to room temperature. Dry by lyophilization. Dissolve 0.1 gof lyophilized powder in 100 g H₂O. Add 10 g of tomato pulp (Lycored).Homogenize suspension. Dry by spray dry method.

Application: Add dried powder to 1270 g soy mass (“Tivall”), homogenize,pack under vacuum in special cover, heat in water (90° C., 15 min.) andstore. The soy mass remains with a pink-red color.

Example 6

Disperse 0.2 g of isolated soy protein 590 in 99 g H₂O. Adjust pH ofsuspension up to 9.2 as per need with 0.5 M NaOH. Heat the dispersion at70° C. for 30 min. Cool to room temperature and dry by lyophilization.Dissolve 0.2 g of lyophilized powder in 100 g H₂O. Add 11 g of tomatopulp (Lycored), homogenize pulp suspension (Silverson machine). Dry pulpwith protein by spray dry method. Application: Add dry powder (11.2 g)to 1270 soy mass (“Tivall”), homogenize, pack under vacuum in specificcover, heat in water (90° C., 15 min.) and store. The soy mass remainswith a pink-red color.

Example 7

Disperse 0.5 g isolated soy protein 590 in 99 g H₂O. Adjust pH ofsuspension to 9.4 with 0.5 M NaOH. Heat the dispersion at 70° C. for 30min. Cool to room temperature. Add 10 g of tomato pulp (Lycored) toprepared solution of water-soluble protein. Homogenize dispersion(Silverson machine) and dry by spray dry method.

Application: Add dry powder (10.5 g) to 1270 g soy mass (“Tivall”),homogenize, pack under vacuum in special cover, heat in water (90° C.,15 min.) and store.

Although the invention has been described in conjunction with specificembodiments, it is evident that many alternatives and variations will beapparent to those skilled in the art in light of the foregoingdescription. Accordingly, the invention is intended to embrace all ofthe alternatives and variations that fall within the spirit and scope ofthe appended claims.

1. In a method for coloring a food, pharmaceutical or cosmetic having afat and/or oil content greater than 5%, with a red coloring material,the improvement wherein said red coloring material is a lycopene coatedwith a water non-soluble thin film comprising an amphiphilic proteinpolymer.
 2. The method according to claim 1, wherein said lycopene isselected from the group consisting of synthetic lycopene, tomato pulp,and lycopene extracted from biomass.
 3. The method according to claim 1,wherein said amphiphilic protein is selected from the group consistingof hydrophobically modified soy proteins, whey protein isolates, eggalbumin, lysozymes, modified pea proteins, gelatin and mixtures thereof.4. The method according to claim 1, wherein said water non-soluble thinfilm further comprises a colloid.
 5. The method according to claim 4,wherein said colloid is selected from the group consisting of gumarabic, xanthan gum, amidated starch, amidated pectins, food gradepolysaccharides, pectins, alginates, xanthan, tragacanth, modifiedpectins, modified alginates, modified xanthan, modified tragacanth,modified starch, moditified chitosans, maltodextrin, modifiedmethylcellulose, galactomannan and mixtures thereof.
 6. A process forthe preparation of stable lycopene formulation comprising (a) treatingan isolated protein to form a protein in a molecular form; (b)dispersing lycopene in an aqueous solution comprising an isolatedprotein in a molecular form; (c) grinding said dispersion to formlycopene particle size of 1 to 10 μm forming an homogenized mixturecomprising fine particles; and optionally (d) drying the homogenizedmixture.
 7. A process according to claim 6, further comprising the stepof mixing said lycopene particles with at least one colloid in waterfollowed by drying.
 8. A process according to claims 6, wherein saidlycopene is selected from the group consisting of synthetic lycopene,tomato pulp, and lycopene extracted from biomass.
 9. A process accordingto claim 6, wherein said isolated protein is amphiphilic and is selectedfrom the group consisting of hydrophobically modified soy proteins, wheyprotein isolates, egg albumin, lysozymes, modified pea proteins, gelatinand mixtures thereof.
 10. The method according to claim 7, wherein saidcolloid is is selected from the group consisting of gum arabic, xanthangum, amidated starch, amidated pectins, food grade polysaccharides,pectins, alginates, xanthan, tragacanth, modified pectins, modifiedalginates, modified xanthan, modified tragacanth, modified starch,moditified chitosans, maltodextrin, modified methylcellulose,galactomannan and mixtures thereof.
 11. A process according to claims 6,wherein said lycopene formulation further comprises an antioxidantand/or emulsifier.
 12. A process according to claim 11, wherein saidantioxidant is ascorbic acid, citric acid, tocopherol or a mixture oftwo or more thereof.
 13. A lycopene containing dry powder obtainable bythe process of claim
 6. 14. A lycopene containing dry powder obtainableby the process of claim
 7. 15. A lycopene containing dry powder asclaimed in claim 13 with a lycopene content between 1% to 15% (w/w). 16.A method for coloring foods, cosmetics or pharmaceuticals having an oilor fat contents higher than 5% with a natural red color, comprising theaddition of a lycopene containing powder of claim
 13. 17. A method forcoloring foods, cosmetics or pharmaceuticals having an oil or fatcontents higher than 5% with a natural red color, comprising theaddition of a lycopene containing powder of claim
 14. 18. A lycopenecoated with a water non-soluble thin film comprising at least onepolymer for use in coloring with red color foods, pharmaceuticals orcosmetics having fat or oil contents higher than 5%.
 19. A coatedlycopene according to claim 18, wherein said lycopene is selected fromthe group consisting of synthetic lycopene, tomato pulp, or lycopeneextracted from biomass.
 20. A coated lycopene according to claim 18,wherein said amphiphilic protein is selected from the group consistingof hydrophobically modified soy proteins, whey protein isolates, bovineserum albumin, caseines, lactoglobulins, egg albumin, lysozymes,modified pea proteins, gelatin and mixtures thereof.
 21. A coatedlycopene according to claim 18, wherein said water non-soluble thin filmfurther comprises a colloid.
 22. The method according to claim 21,wherein said colloid is selected from the group consisting of gumarabic, xanthan gum, amidated starch, amidated pectins, food gradepolysaccharides, pectins, alginates, xanthan, tragacanth, modifiedpectins, modified alginates, modified xanthan, modified tragacanth,modified starch, moditified chitosans, maltodextrin, modifiedmethylcellulose, galactomannan and mixtures thereof.
 23. The dry powderof claim 15 having a lycopene content of 4-8%.
 24. The coated lycopeneof claim 19, wherein the lycopene is extracted from tomato pulp.
 25. Theprocess of claim 8, wherein the lycopene is extracted from tomato pulp.26. The method of claim 2, wherein the lycopene is extracted from tomatopulp.